AU747285B2 - Antimicrobial composition - Google Patents

Description

S F Ref: 433155

AUSTRALIA

PATENTS ACT 1990 COMPLETE SPECIFICATION FOR A STANDARD PATENT

ORIGINAL

r Name and Address of Applicant: Actual Inventor(s): Address for Service: F. Hoffmann-La Roche AG 124 Grenzacherstrasse CH-4070 Basle

SWITZERLAND

Peter Angehrn, Ingrid Heinze-Krauss, Malcolm Page, Hans G.F. Richter Spruson Ferguson, Patent Attorneys Level 33 St Martins Tower, 31 Market Street Sydney, New South Wales, 2000, Australia Invention Title: Antimicrobial Composition The following statement is a full description of this invention, including the best method of performing it known to me/us:- 5845 The present invention relates to antimicrobial compositions comprising a combination ofvinyl-pyrrolidinone cephalosporin derivatives with carbapenem antibiotics or with p-lactamase inhibitors, to the use of the compositions for producing a medicament for the treatment and prophylaxis of infectious diseases, including MRSA infections, and to pharmaceutical compositions containing the combination.

The extensive use of -lactam antibiotics during the last decades result in a considerable increase of pathogens resistant to this class of antibiotics. A major threat are infections caused by Methicillin resistant Staphylococcus aureus (MRSA). Various strains of MRSA have been reported to be resistant not only to P-lactams but also to many other antimicrobial agents, including aminoglycosides, erythromycin, chloramphenicol and quinolones.

Vinyl-pyrrolidinone cephalosporin derivatives and their preparation are described in the European Patent Application 96117710.2 filed January 10, 1997. These compounds exhibit a high antimicrobial activity against non- P-lactamase producing strains of MRSA. In P-lactamase-producing strains of MRSA partial hydrolysis of vinyl-pyrrolidinone cephalosporins can occur.

It has now been found that a combination of vinyl-pyrrolidinone cephalosporins of the formula I with p-lactamase inhibitors like, e.g.

clavulanic acid, Tazobactam, Sulbactam or (Z)-(2S,3S,5R)-3-(2-cyanoethenyl)- 3-methyl-4,4,7-trioxo-4-thia-l-aza-bicyclo[3.2.0]heptane-2-carboxylic acid a compound as described in the European Patent Application EP-A-0 640 607) prevents these compounds from being hydrolyzed by staphylococcal 1lactamase thus, resulting in a synergistic effect.

Carbapenems like, e.g. Imipenem.or Meropenem are p-lactam antibiotics that have a broad spectrum of antibacterial activity and are in addition good P-lactamase inhibitors. However they do not cover MRSA.

It has now been found that a combination of vinyl-pyrrolidinone cephalosporins of the formula I with carbapenems also leads to a synergistic effect.

Compared to known combinations of two P-lactam antibiotics as described in the European Patent EP-A-384410, Banyu Pharmaceutical Co.

Ltd; and compared to a combination of cephalosporins such as e.g. ceftriaxone and meropenem as described in the Japanese Laid Open Patent Publication JP-02-279627 A, Sumitomo, the combinations of the present invention show a higher efficacy against MRSA.

Furthermore, the combination comprises a unique broad spectrum of antibacterial activity that includes Gram-positive pathogens like, e.g.

methicillin-sensitive and methicillin-resistant staphylococci, enterococci, penicillin-sensitive and penicillin-resistant pneumococci and Gram negative pathogens like, e.g. E. coli, E. cloacae, K. pneumoniae, P. vulgaris, S.

marcescens, C. freundii and P. aeruginosa.

Accordingly, the present invention provides an antimicrobial composition comprising a combination of 15 a first ingredient selected from vinyl-pyrrolidinone cephalosporin derivatives of the general formula I

H

R 1-(Xs-(CR 4 R 5)m N S I

(CH

2 )n

COR

3 wherein

R

1 is lower alkyl, halogen, phenyl, benzyl, styryl, naphthyl or 20 heterocyclyl; the lower alkyl, phenyl, benzyl, styryl, naphthyl and heterocyclyl being optionally substituted by at least one of halogen, hydroxy, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted phenyl, amino, lower alkylamino, di-lower alkylamino, carboxy, lower alkylcarboxy, carbamoyl or lower alkylcarbamoyl;

R

4

R

5 independently are hydrogen, lower alkyl or phenyl; X is S, O, NH or CH 2 n is 0, 1 or 2; m is 0 or 1; s is 0 or 1;

R

2 is hydrogen, hydroxy, carbamoylmethyl, -CH2-CO-NH-R 6 lower alkyl-Qr, cycloalkyl-Qr, lower alkoxy, lower alkenyl, cycloalkenyl-Qr, lower alkynyl, aralkyl-Qr, aryl-Qr, aryloxy, aralkoxy, heterocyclyl or heterocyclyl-Qr; the lower alkyl-, cycloalkyl-, lower alkoxy-, lower alkenyl-, cycloalkenyl-, lower alkynyl-, aralkyl-, aryl-, aryloxy-, aralkoxyand the heterocyclyl-group may be substituted with at least one group selected from carboxy, amino, nitro, cyano, -S02-NH-R 6 optionally fluoro substituted lower alkyl, lower alkoxy, hydroxy, halogen, -CONR 6

R

7 -CH2-CONR 6

R

7

N(R

7

)COOR

8

R

7

CO-,

R

7 OCO-, R 7 COO-, -C(R 7

R

9 )CO2R 8

-C(R

7

R

9

)CONR

7

R

1 0 wherein

R

6 is hydrogen, lower alkyl, cycloalkyl or aryl;

R

7 and R 9 are independently hydrogen or lower alkyl; 15 R 8 is hydrogen, lower alkyl, lower alkenyl or a carboxylic acid protecting group; and

R

10 is hydrogen, o-hydroxy-alkyl, phenyl, naphthyl or heterocyclyl; the phenyl, naphthyl or heterocyclyl being unsubstituted or 20 substituted with at least one of the groups of optionally protected hydroxy, halogen, optionally substituted lower alkyl or o-hydroxyalkyl, optionally substituted lower alkoxy and/or cyano; or R 7 and R 10 form together a group of formula "25 Q is -CH2-, -CH-lower-alkyl, -CO- or -SO 2 r is 0 or 1; and

R

3 is hydroxy, or -OM and M represents an alkali metal; as well as readily hydrolyzable esters thereof, pharmaceutically acceptable salts of said compounds and hydrates of the compounds of formula I and of their esters and salts; and a second ingredient selected from the class of carbapenem antibiotics and pharmaceutically acceptable salts thereof or from 3-lactamase inhibitors and pharmaceutically acceptable salts thereof.

The term "carbapenem antibiotics" denotes compounds containing the structural element 0

H

and exerting an antibiotic activity, such as e.g. imipenem or meropenem.

Imipenem is preferred.

Preferred "P-lactamase inhibitors" are tazobactam, sulbactam,.clavulanic acid or 3 -0-alkenyl-penam-sulfon compounds as described in the European Patent EP-0 640 607 B1, particularly 2 S,3S,5R)-3-(2-cyanoethenyl)-3methyl-4,4,7-trioxo-4-thia-l-aza-bicyclo[3.2.0]heptane-2-carboxylic acid and pharmaceutically acceptable salts having the following formula

O

S OH 3

CN

N

O

COOH

10 The compositions of the present invention are effective against MRSA which are highly resistant to 3-lactam antibiotics and furthermore, provide a synergistic effect as compared to the compounds used alone.

As used herein pharmaceutically acceptable salts useful in this invention include salts derived from metals, the ammonium salt, quaternary ammonium salts derived from organic bases, amines and amino acid salts. Examples of preferred metal salts are those derived from the alkali metals, for example, lithium sodium and potassium Examples of quaternary ammonium salts derived from organic bases include tetramethylammonium (N+(CH3) 4 tetraethylammonium (N+(CH2CH 3 4 S 20 benzyltrimethylammonium (N+(C6H5CH2)(CH 3 3 phenyltriethylammonium (N+(C6H5)(CH 2

CH

3 3 and the like, etc. Those salts derived from amines include salts with N-ethylpiperidine, procaine, lidocaine, dibenzylamine,

N,N'-

dibenzylethylenediamine, alkylamines or dialkylamines as well as salts with amino acids such as, for example, salts with arginine or lysine. With respect to vinyl-pyrrolidinone derivatives having basic or quaternary substituents

R

2 also salts with mineral or organic acids are included. Especially preferred are hydrochlorides, sulfates, phosphates, lactates, mesylates or the inner salt.

A second aspect of the present invention provides a pharmaceutical preparation containing an antimicrobial composition comprising a first ingredient and a second ingredient as defined above together with a therapeutically inert carrier.

A third aspect of the present invention provides a method for the treatment or prophylaxis of infectious diseases in a mammal requiring said treatment or prophylaxis, which method includes or consists of administering to said mammal an effective amount of at least one antimicrobial composition of the invention as defined above or of a pharmaceutical preparation of the second aspect of the invention.

A fourth aspect of the present invention provides an antimicrobial composition of the present invention as defined above, or of a pharmaceutical preparation of the second aspect of the invention, when used in the treatment or phrophylaxis of infectious diseases.

15 A fifth aspect of the present invention provides the use of an antimicrobial :composition of the present invention as defined above for the manufacture of a medicament for the treatment or prophylaxis of infectious diseases.

As readily hydrolyzable esters of the compounds of formula I there are to be aunderstood compounds of formula I, the carboxy group(s) of which (for example, the 2 -carboxy-group) is/are present in the form of readily •oo a a. 0 a hydrolyzable ester groups. Examples of such esters, which can be of the conventional type, are the lower alkanoyloxy-alkyl esters the acetoxymethyl, pivaloyloxymethyl, 1-acetoxyethyl and 1-pivaloyloxyethyl ester), the lower alkoxycarbonyloxyalkyl esters the methoxycarbonyloxymethyl, 1-ethoxycarbonyloxyethyl and 1-isopropoxycarbonyloxyethyl ester), the lactonyl esters the phthalidyl and thiophthalidyl ester), the lower alkoxymethyl esters the methoxymethyl ester) and the lower alkanoylaminomethyl esters the acetamidomethyl ester). Other esters the benzyl and cyanomethyl esters) can also be used. Other examples of such esters are the following: (2,2-dimethyl-l-oxopropoxy)methyl ester; methylpropoxy)carbonyl]-2-pentenyl ester; 1- [[(1-methylethoxy)carbonyl]oxy] ethyl ester; 1-(acetyloxy) ethyl ester; (5-methyl-2-oxo-l,3-dioxol-4-yl) methyl ester; 1-[[(cyclohexyloxy)carbonyl]oxy] ethyl ester; and 3,3-dimethyl-2-oxobutyl ester. It will be appreciated by those of ordinary skill in the art that the readily hydrolyzable esters of the compounds of the present invention can be formed at a free carboxy group of the compound.

The compounds of formula I as well as their salts and readily hydrolyzable esters can be hydrated. The hydration can be effected in the course of the manufacturing process or can occur gradually as a result of hygroscopic properties of an initially anhydrous product.

Examples of salts of the compounds of formula I are defined under "pharmaceutically acceptable salts" above.

In above compounds of formula I the substituent in position 3 can be present in the E-form [Ia] or in the Z-form [Ib].

(I R 2 1001 N-R 2 Ia Ib 0 Compounds of formula Ia i.e. wherein the substituent in position 3 is in the Eform are generally preferred.

In a particular embodiment of the compounds of formula I n is 1.

The term "halogen" or "halo" used herein refers to all four forms, that is chlorine or chloro; bromine or bromo; iodine or iodo; and fluorine or fluoro, unless specified otherwise.

-6- As used herein, the terms "alkyl" and "lower alkyl" refer to both straight and branched chain saturated hydrocarbon groups having 1 to 8, and preferably 1 to 4, carbon atoms, for example, methyl, ethyl, n-propyl, isopropyl, t-butyl and the like.

By the term "optionally substituted lower alkyl" is meant a "lower alkyl" moiety as defined above substituted by, for example, halogen, amino, hydroxy, cyano, carboxy, carbamoyl, etc., such as carboxymethyl, 2-fluoroacetyl, trifluoromethyl, 2,2,2-trifluoroethyl, 2-chloroethyl, hydroxymethyl, 2hydroxyethyl, carbamoylmethyl and like.

As used herein, the term "lower alkoxy" refers to a straight or branched chain hydrocarbonoxy group wherein the "alkyl" portion is a lower alkyl group as defined above. Examples include methoxy, ethoxy, n-propoxy and the like.

The "alkyl" portion may be substituted as defined above.

As used herein, "alkenyl" and "lower alkenyl" refer to unsubstituted or 15 substituted hydrocarbon chain radical having from 2 to 8 carbon atoms, preferably from 2 to 4 carbon atoms, and having at least one olefinic double bond, e.g. allyl, vinyl etc.

As used herein, "lower alkynyl" refers to unsubstituted or substituted hydrocarbon chain radical having from 2 to 8 carbon atoms, preferably 2 to 4 carbon atoms, and having at least one triple bond.

By the term "cycloalkyl" is meant a 3-7 membered saturated carbocyclic moiety, cyclopropyl, cyclobutyl, cyclohexyl, etc.

As used herein, "cycloalkenyl" refers to a carbocyclic ring radical having at least one olefinic double bond.

By the term "aryl" is meant a radical derived from an aromatic hydrocarbon by the elimination of one atom of hydrogen and can be substituted or unsubstituted. The aromatic hydrocarbon can be mononuclear or polynuclear. Examples of aryl of the mononuclear type include phenyl, tolyl, xylyl, mesityl, cumenyl, and the like. Examples of aryl of the polynuclear type include naphthyl, anthranyl, phenanthryl, and the like. The aryl group can have at least one substituent selected from, as for example, halogen, hydroxy, cyano, carboxy, nitro, amino, lower alkyl, lower alkoxy, carbamoyl, sulfamoyl; such as in 2,4-difluorophenyl, 4-carboxyphenyl, 4-nitrophenyl, 4-aminophenyl, -7- 4-methoxyphenyl, 3-hydroxyphenyl, 4-hydroxyphenyl, 3-fluor-4hydroxyphenyl, 4-sulfamoylbenzyl.

By the term "aralkyl" is meant an alkyl group containing an aryl group.

It is a hydrocarbon group having both aromatic and aliphatic structures, that is, a hydrocarbon group in which a lower alkyl hydrogen atom is substituted by a monocyclic aryl group, phenyl, tolyl, etc. Such groups are e.g. benzylgroups.

As used herein, "aryloxy" is an oxygen radical having an aryl substituent -O-aryl).

As used herein, "aralkoxy" is an oxygen radical having an aralkyl substituent.

As used herein, the term "lower alkylamino and di-lower alkylamino" refers to mono and dialkylamino residues wherein lower alkyl is as defined above, for example methylamino, 2-ethylamino, -CH 2

NHCH

3

-CH

2

CH

2

NHCH

3

-CH

2

CH

2 N(CH3) 2 N-methylamino, N-ethylamino, N,Ndimethylamino, N,N-diethylamino and the like.

As used herein, "heterocyclic ring" or "heterocyclyl"refers to an unsaturated or saturated, unsubstituted or substituted, mono or bicyclic or 7-membered heterocyclic ring containing at least one hetero atom selected from the group consisting of oxygen, nitrogen, or sulfur. Exemplary heterocyclic rings include, but are not limited to, for example, the following groups: azetidinyl, pyridyl, pyrazinyl, piperidyl, piperidino, N-oxido-pyridyl, pyrimidyl, piperazinyl, pyrrolidinyl, pyridazinyl, N-oxide-pyridazinyl, pyrazolyl, triazinyl, imidazolyl, thiazolyl, quinoline, benzothiazolyl, benzoxazolyl, benzimidazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4thiadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3triazolyl, 1,2,4-triazolyl, 1H-tetrazolyl, 2H-tetrazolyl; furyl, 1H-azepinyl, thiophenyl, tetrahydrothiophenyl, tetrahydrofuranyl, isoxazolyl, isothiazolyl, oxazolidinyl, 1H-pyrazolo[3,4-d pyrimidin-4-yl, [1,2,4]triazololl 7-yl, 5-oxo-2,5-dihydro-[1,2,4]triazin-3-yl, 2-oxo-3,7-dihydro-2H-purin-6-yl, etc.

Substituents for the heterocyclic ring include, for example, lower alkyls such as methyl, ethyl, propyl, etc., lower alkoxys such as methoxy, ethoxy, etc., halogens such as fluorine, chlorine, bromine, etc., halogen substituted alkyls such as trifluoromethyl, trichloroethyl, etc., amino, mercapto, hydroxyl, carbamoyl, or carboxyl groups. A further substituent is oxo, such as in 2-oxo- -8oxazolidin-3-yl, 1,1-dioxo-tetrahydrothiophen-3-yl. Further examples of substituted heterocycles are 6-methoxy-pyridin-3-yl, 5-methyl-isoxazol-3-yl, 1methyl-pyridinium-2-yl, -3-yl, 4 -yl, 1-carbamoylmethyl-pyridinium-2-yl, 1-carbamoylmethyl-pyridinium-3-yl, 1-carbamoylmethyl-pyridinium-2-yl, -3yl, -4-yl, 1-[N-(4-hydroxy)phenyll-carbamoylmethyl-pyridinium-4-yl, fluoro-4-hydroxy)phenyl] -carbamoylmethyl-pyridinium-4-yl.

By the term "substituted phenyl" is meant phenyl mono or di-substituted by halogen, optionally substituted lower alkyl, optionally protected hydroxy or amino, nitro or trifluoromethyl.

The term "optionally protected hydroxy refers to hydroxy or hydroxy protected, for example with t-butyloxycarbonyl, trimethylsilyl, t-butyldimethylsilyl, tetrahydropyranyl, trifluoroacetyl, and the like, or refers to an ester group, for example, phosphate or sulfonate.

The term "optionally protected amino" refers to amino or amino protected with, for example, BOC [t-butoxycarbonyl; other name: (1,1-dimethylethoxy)carbonyl], benzyloxycarbonyl and allyloxycarbonyl.

The term "carboxylic acid protecting group" refers to protecting groups conventionally used to replace the acidic proton of a carboxylic acid.

Examples of such groups are described in Greene, Protective Groups 20 in Organic Synthesis, Chapter 5, pp. 152-192 (John Wiley and Sons, Inc.

1981), incorporated herein by reference. Preferably these examples include methoxymethyl, methylthiomethyl, 2,2,2-trichloroethyl, 2-haloethyl, 2- (trimethylsilyl)ethyl, t-butyl, allyl, benzyl, triphenylmethyl (trityl), benzhydryl, p-nitrobenzyl, p-methoxybenzyl, trimethylsilyl, triethylsilyl, tbutyldimethylsilyl, i-propyl-dimethylsilyl. Preferred are benzyhydryl, t-butyl, p-nitrobenzyl, p-methoxybenzyl and allyl.

A preferred embodiment of the invention are compounds of formula I wherein R1 is phenyl, 3,4-dichlorophenyl, 2,5-dichlorophenyl, dichlorophenyl, 2,4,5-trichlorophenyl, 4-trifluoromethyl-phenyl, 4-methoxyphenyl, 4-hydroxymethylphenyl, 3,4dimethoxyphenyl, 2-naphthyl, benzyl, 2-benzothiazolyl, 2benzoxazolyl, 4-methyl-1,2,4-triazol-5-yl, 9benzimidazol-2-yl, (2-amino)-thiazol-4-yl, pyridin-4-yl, pyrimidin-2-yl, pyridinium-1-yl, 2-yl, -3-yl or -4-yl; is cyclopropyl, cyclopropylmethyl, 1,1,1-trifluorethyl, or 4hydroxybenzyl, 4-hydroxyphenyl, 4-sulfamoylbenzyl, pyrrolidin- 3-yl or a group of formula wherein Q1 is -CH 2 r 10 R1 x

(CR

4

R

5 )m s is 0 or 1; is hydrogen, lower alkyl, co-hydroxy alkyl, benzyl or alkylheterocyclyl; the benzyl and the heterocyclyl group being unsubstituted or substituted with at least one of the groups cyano, carboxy or hydroxy; or is -CH 2

CONR

7 R1 0 wherein R 7 and R 10 are as defined above.

is hydroxy, or -OM and M represents an alkali metal; is S; is -His 1; and pharmaceutically acceptable salts thereof.

An especially preferred embodiment of the invention are compounds of formula I wherein

R

is phenyl, 2-naphthyl, 2,5-dichlorophenyl, 3 2,4,5-trichlorophenyl, 2-benzothiazolyl, 4-pyridinyl; is cyclopropyl, methyl-cyclopropyl, 3- or 4-hydroxybenzyl, 4hydroxyphenyl, 4-sulfamoylbenzyl, pyrrolidin-3-yl, 1- hydroxy)phenyl] -carbarnoylmethyl-pyridin- 1-ium-4-yl, 1- fluoro-4-hydroxy)phenyll -carbamoylmethyl-pyridin- 1-ium-4-yl.

is hydroxy, lower-alkoxy, or -OM and M represents an alkali metal; is S; is -CH 2 x

(CR

4

R

5 )m s isi1; and pharmaceutically acceptable salts thereof.

Especially preferred compounds of formula I are: [1-11-[(4-Hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1-ium-4ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino] -5-thia- 1-aza-bicyclo oct-2-ene-2-carboxylate (Example 1), 4 Q /OH C

HH

C1 0 [1-[(4-Hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1-ium-4ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyll -8-oxo-7- 1-benzothiazol-2-ylsulfanyl)-acetylamino] -5-thia- 1-aza-bicyclo [4.2 .0]oct-2-ene-2-carboxylate ::*(Example 2), .9 9*H N 0 *C02- 0 [2-(Benzothiazol-2-ylsulfanyl)-acetylaminol 1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia-1-aza-bicyclo oct- 2-ene-2-carboxylic acid (Example 3), Q

HN

CO 2 H- 0 [(4-Hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1-ium-4ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-7- [2-(pyridin-4-ylsulfanyl)acetylamino] -5-thia- 1-aza-bicyclo oct-2-ene-2-carboxylate (Example 4), 11

OH

N 0

H

[1-l-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1ium-4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyl] [2-(naphthalen-2-ylsulfanyl)-acetylamino] -8-oxo-5-thia-1-aza-bicyclo oct-2-ene-2-carboxylate (Example

S

S

S S S. S

SS*

S

[1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylaminol -5-thia- 1-aza-bicyclo oct- 2-ene-2-carboxylic acid (Example 6), CI CI H

~OH

10 COOH 0 (E)-(6R,7S)-3-[Il-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-7- [2-naphthalen-2-ylsulfanyl)-acetylamino -5-thia- 1-aza-bicyclo oct-2-ene- 2-carboxylic acid (Example 7),

H

0 N aOH C0 2 H 0 Mixture of and -[(S)-2-oxo-[1,3']Bipyrrolidinyl-3-ylidenemethyl] -8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino] -5-thia- 1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid hydrochloride (Example 8), 12 1j N1 1*HCI C0 2 H 0 [1-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1ium-4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-7-(2-phenylsulfanyl- 1-aza-bicyclo oct-2-ene-2-carboxylate (Example 9) 4 Nlr-

OH

S. S

S.

S S *5 e S. S S S ,5-Dichloro-phenylsulfanyl)-acetylamino] [1-[(3-fluoro-4hydroxy-phenylcarbamoyl)-methyll -pyridin- 1-ium-4-ylmethyl] -2-oxopyrrolidin-3-ylidenemethyl] -8-oxo-5-thia- 1-aza-bicyclo [4.2.01 oct-2-ene-2carboxylate (Example (E)-(6R,7R)-8-oxo-3- [1 ,3']Bipyrrolidinyl-3-ylidenemethyl] [2- (phenylsulfanyl)-acetylamino] -5-thia- 1-aza-bicyclo oct-2-ene-2-carboxylic acid hydrochloride (Example 11),

HC

-13- 1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7-[2- (naphthalene-2-ylsulfanyl)-acetylamino] -8-oxo-5-thia-1-aza-bicyclo[4.2.0]oct-2ene-2-carboxylic acid (Example 12),

H

N s 0

CO

2 H O (E)-(6R,7R)-3-[1-(4-Hydroxy-phenyl)-2-oxo-pyrrolidin-3-ylidenemethyl -7- [2-(naphthalene-2-ylsulfanyl)-acetylamino] -8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid sodium salt (Example 13),

H

NN

OH

o

CO

2 Na O (E)-(6R,7R)-8-Oxo-3-[2-oxo- 1-(4-sulfamoyl-benzyl)-pyrrolidin-3- 10 ylidenemethyl] -7-[2-phenylsulfanyl)-acetylamino] -5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (Example 14).

SO

2

NH

2

N

0 see

CO

2 H O In one aspect of the invention the first ingredient and the second ingredient as defined above are formulated in a single composition, and in another embodiment, the first and the second ingredient are separately formulated in different compositions. Thus, the invention also comprises a kit consisting of a composition containing a compound of formula I in dosage unit form and a second composition in dosage unit form containing a carbapenem antibiotic or a P-lactamase inhibitor as defined earlier.

The ratio of the first to the second ingredient can be widely varied, and is usually about 1:20 to about 20:1.

The present composition can be administered in the same manner as the ingredients alone, can be administered parenterally, orally or topically.

-14- For parenteral administration the compositions are preferably made into preparations as lyophilisates or dry powders for dilution with customary agents, such as water, isotonic common salt solution or carbohydrate.

Where the composition comprising both the first and the second ingredient is administered the total amount administered is 0,25 8 g/day.

The synergism of the present composition is demonstrated by testing representative compounds of the present invention. In vitro activity was determined by minimum inhibitory concentration (MIC) by the agar dilution method in Mueller Hinton agar inoculum 10 5 CFU/spot.

Table 1 shows the in vitro activity (MIC, of selected cephalosporins in combination with 4 mg/L Imipenem against 17 highly resistant MRSA strains.

9* Table 1 555555 0

SS

S

S.

S.

S 5 5*

S

Example MIC 50

MIC

90

MIC

100 range of MIC Imipenem >16 >16 >16 16->16 Meropenem >16 >16 >16 16->16 Cefoperazone >64 >64 >64 >64 Cefotiam >64 >64 >64 >64 Cefpiramid >64 >64 >64 >64 Ceftriaxone >64 >64 >64 >64 Cefotaxime >64 >64 >64 >64 Cefoperazone Imipeneml) >64 >64 >64 >64 Cefotiam Imipeneml) >64 >64 >64 64- >64 Cefpiramid Imipenem 1 >64 >64 >64 32 >64 Ceftriaxone Imipenem >64 >64 >64 >64 Cefotaxime Imipenem >64 >64 >64 >64 Ceftriaxone Meropenem 2 >64 >64 >64 >64 Cefotaxime Meropenem 2 >64 >64 >64 >64 Ex.1 >16 >16 >16 2->16 Ex.1 Imipenem 0.5 1 2 <0.12 2 Ex.2 >16 >16 >16 2->16 Ex.2 Imipenem 1 1 2 0.25 2 Ex.3 16 >16 >16 2->16 Ex.3 Imipenem 2 4 4 1-4 Ex.4 >16 >16 >16 4- >16 Ex.4 Imipenem 1 2 2 0.25 2 Ex.5 >16 >16 >16 2 >16 Imipenem 0.5 2 2 <0.12 2 Ex.6 16 >16 >16 1->16 Ex.6 Imipenem 2 4 4 <0.12 4 Ex.7 16 16 16 2-16 Ex.7+ Imipenem 1 4 4 0.5-4 Ex.8 >16 >16 >16 2 >16 Ex.8 Imipenem 0.5 2 4 <0.12 4 Ex.9 16 16 >16 1->16 Ex.9 Imipenem 0.5 1 1 <0.12 1 >16 >16 >16 1->16 Imipenem 0.5 1 2 <0.12 2 0 0050 16 Example MIC 5 0 MIC 9 0 MI~ioo range of MIC Ex.11 16 >16 >16 4 ->16 Ex. 11 Imipenem 0.5 1 2 <0.25 2 Ex.12 16 >16 >16 4 ->16 Ex.12 Imipenem 2 4 4 0,5 -4 Ex. 13 16 >16 >16 2- >16 Ex. 13 Imipenem 1 2 2 <0.25 2 Ex. 14 16 >16 >16 1- >16 Ex. 14 Imipenem 1 14 18 <0.25 8 1) combination described in EP-A 0 384 410 2) combination described in JP-02-279627 A Table 2 shows the in vitro activity (MIC, [mg/LI) of selected cephalosporins in combination with 4 mg/L compound B (sodium cyanoethenyl)-3-methyl-4,4,7-trioxo-4-thia- 1-aza-bicyclo 13.2 .0]heptane-2carboxylate) of the formula against 17 highly resistant MRSA strains -17- Table 2 Example MIC 50

MIC

90 M1C 100 range of MIC compound B >16 >16 >16 ->16 Ex.1 >16 >16 >16 2 ->16 Ex. 1+ compound B 2 2 2 1-2 Ex.2 >16 >16 >16 2 >16 Ex.2+ compound B 2 2 4 2-4 Ex.4 >16 >16 >16 4 >16 Ex.4 +compound B 4 8 8 4-8 >16 >16 >16 2 >16 +compound B 2 2 2 1-2 Ex.6 16 >16 >16 1 >16 Ex.6 +compound B 2 2 4 1-4 Ex.7 16 16 16 2-16 Ex.7 +compound B 2 4 4 2-4 Ex.8 >16 >16 >16 2 >16 Ex.8 +compound B 2 4 4 0.5-4 Ex.9 16 16 >16 1 ->16 Ex.9 +compound B 2 2 2 0.5-2 >16 >16 >16 1 >16 compound B 1 2 2 0.5-2 Ex.11 16 >16 >16 4 ->16 Ex. 11 +compound B 2 2 4 1-4 Ex.12 16 >16 >16 4 >16 Ex. 12 +compound B 4 8 8 2-8 The compounds of the formula I as well as their pharmaceutical acceptable salts, hydrates, or readily hydrolyzable esters can be manufactured by treating a compound having the formula II R'HN S(CH2)n 0CH

N-R

2 COOR 9 0 in which -18-

R

2 and n are defined above;

R

f is hydrogen or trimethylsilyl; and Rg is hydrogen, benzhydryl, p-methoxybenzyl, t-butyl, trimethylsilyl or allyl or salt thereof, with a carboxylic acid of the general formula III

R

1 s-(CR 4

R

5 0 III in which

R

1 X, s, R 4

R

5 and m are as defined above, and Y is -OH, or a reactive functional derivative thereof wherein Y is, for example 10 a halogen as chloride or bromide, or 1-imidazolyl, 2-mercaptobenzotriazolyl, 1hydroxy benzotriazolyl or pivaloyloxy; for compounds of formula I wherein X is S, 0 or NH, by treating a compound having the formula IV

H

N S (CH 2 )n 0 N N-R 2 CO2R O IV 0 wherein R 4

R

5 m, n, R 2 and Rg are as defined above and Hal stands for halogen as bromine or chlorine or iodide preferably bromine, with an appropriate thiol or thiolate or an appropriate alcohol or alcoholate or an appropriate amine in the presence of a base, for the manufacture of a readily hydrolysable ester of a compound of formula I subjecting a carboxylic acid of formula I to a corresponding esterification or for the manufacture of salt or hydrates of a compound of formula I or hydrates of said salts converting a compound of formula I into a salt or hydrate or into a hydrate of said salts.

-19- The reaction of a compound of formula II prepared according to embodiment with a compound of formula III, or a reactive derivative thereof can be carried out in a manner known per se. The carboxy groups in compounds of formula II (carboxy group in position 2 and/or carboxy groups optionally present in R 2 in compounds of formula III (carboxy groups optionally present in R 1 can be protected intermediatly or in situ, for example, by esterification to form readily cleavable esters such as a silyl ester trimethylsilylester), a p-methoxy-benzylester or benzhydryl ester.

Furthermore the amino groups present in compounds of formula II (in position 7 and/or optionally present in R 2 and/or optionally present in R 1 of compounds of formula III can be protected, for example, with protecting groups which are cleavable by acid hydrolysis the t-butoxycarbonyl or triphenylmethyl groups), by basic hydrolysis the trifluoroacetyl group), by hydrazinolysis the phthalimido group) or by catalytic cleavage in 15 presence of Pd (the allyloxycarbonyl group). Preferred protecting groups are the t-butyloxy-carbonyl, allyloxycarbonyl, the chloroacetyl, bromoacetyl and iodoacetyl groups, especially the chloroacetyl group. These last-mentioned protecting groups can be cleaved off by treatment with thiourea. Another preferred protecting group is phenylacetyl which can be cleaved off by treatment with PCl5 or enzymatically.

The 7-amino group in compounds II can be protected in situ, for example, by a silyl protecting group such as the trimethylsilyl group.

In reacting a 7-amino compound of formula II with a carboxylic acid of formula III in the presence of an activating agent such as CDI (1,1'-carbonyldiimidazole), CDT (1,1'-carbonyl-1,2,4-ditriazole), DCC (N,N'-dicyclohexylcarbodiimide), EEDQ (1-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline), HOBT (1-Hydroxybenzotriazole), HBTU (ortho-benzotriazol-1-yl-N,N,N',N'tetramethyluronium-hexafluorphosphate), isobutyl chlorocarbonate, IIDQ (1isobutyloxycarbonyl-2-isobutyloxy-1,2-dihydroquinoline), pivaloyl chloride and the like or a reactive functional derivative of formula III, in an inert solvent such as ethyl acetate, acetonitrile, dioxane, chloroform, methylene chloride, benzene, dimethylformamide or dimethylacetamide can be used. Subsequently the ester group can be cleaved off.

Prepared according to another embodiment, a salt of an acid of formula II a trialkylammonium salt such as the triethylammonium salt) is reacted with a reactive functional derivative of a carboxylic acid of formula III in an inert solvent dimethylformamide, dimethylacetamide, dimethylsulfoxide and the like).

The reaction of a 7-amino compound of formula II with the carboxylic acid of formula III or a reactive derivative thereof can conveniently be carried out at a temperature between about -40 0 C and +60°C, e.g. at room temperature.

Embodiment involves treating a compound of formula IV with an appropriate thiol or thiolate or an appropriate alcohol or alcoholate or an appropriate amine in presence of a base, for example, trialkylamine such as trimethylamine, triethylamine, sodium bicarbonate, DBU (1,8diazabicyclo[5,4,0]undec-7-en(1,5-5)) to form the corresponding thioether, ether or amine. Optionally present amino, hydroxy or carboxyl groups can be intermediately protected with groups as described above.

Deprotection of protected amino, hydroxy or carboxylic groups present in 15 a compound of formulae II, III and IV can be carried out as follows: Removal of amino protecting groups Possible amino-protecting groups are those employed in peptide chemistry, such as the protecting groups mentioned above. Preferably these examples include carbamates, e.g. fluorenylmethyl, 2,2,2-trichloroethyl, tbutyl, triphenylmethyl, allyl, benzyl. Further protecting groups are p-nitro- :benzyl, diphenylmethyl, triphenylmethyl, benzyl, formyl, trifluoroacetyl, S. chloro-acetyl, the cyclic imides of N-phthaloyl, N-trimethylsilyl, Nbenzenesulfonyl, N-toluenesulfonyl. Preferred is BOC [t-butoxycarbonyl; other name: (1,1-dimethylethoxy)carbonyl], benzyloxycarbonyl, allyloxy-carbonyl or 25 trimethylsilyl.

The amino protecting groups may be cleaved off by acid hydrolysis (e.g.

the t-butoxycarbonyl or triphenylmethyl group), e.g. aqueous formic acid, trifluoroacetic acid or by basic hydrolysis the trifluoroacetyl group). The chloroacetyl, bromoacetyl and iodoacetyl groups are cleaved off by treatment with thiourea. The trimethylsilyl group is cleaved off by hydrolysis or alcoholysis.

Amino-protecting groups which are cleavable by acid hydrolysis are preferably removed with the aid of a lower alkanecarboxylic acid which may be halogenated. In particular, formic acid or trifluoroacetic acid is used. The -21reaction is carried out in the acid or in the presence of a co-solvent such as a halogenated lower alkane, e.g. methylene chloride. The acid hydrolysis is generally carried out at room temperature, although it can be carried out at a slightly higher or slightly lower temperature a temperature in the range of about -30°C to Protecting groups which are cleavable under basic conditions are generally hydrolyzed with dilute aqueous caustic alkali at 0 C to 30°C. The chloroacetyl, bromoacetyl and iodoacetyl protecting groups can be cleaved off using thiourea in acidic, neutral or alkaline medium at about 0°C- The phthalimido group can be cleaved off with hydrazine at -20*C to +50 0

C.

Removal of hydroxy protecting groups Possible hydroxy protecting groups are such as are commonly known in the art, such as trimethylsilyl, t-butyl-dimethylsilyl, dimethylphenylsilyl, triphenylmethyl, lower alkanoyl, acetyl, trifluoroacetyl, tetrahydropyranyl, 15 benzyl, p-nitrobenzyl or t-butoxycarbonyl. These groups are removed in the presence of acidic solvents, weak organic acids or weak inorganic bases like sodium bicarbonate, respectively.

Removal of protecting groups at the carboxv function As carboxyl protecting groups one may utilize an ester form which can be easily converted into a free carboxyl group under mild conditions, for example, benzhydryl, t-butyl, p-nitrobenzyl, p-methoxybenzyl, allyl, etc.

These protecting groups may be removed as follows: benzhydryl trifluoroacetic acid with anisol, phenol, cresol or triethylsilane at about -40°C to room temperature; hydrogen with Pd/C in an alcohol such as ethanol or in tetrahydrofuran;

BF

3 -etherate in acetic acid at about 0 to t-butyl formic acid or trifluoroacetic acid with or without anisol, phenol, cresol or triethylsilane and a solvent such as dichloromethane at about -10°C to room temperature; p-nitrobenzyl sodium sulfide in acetone/water at about 0 to room temperature; or hydrogen with Pd/C in an alcohol such as ethanol or in tetrahydrofuran; -22p-methoxybenzyl formic acid at about 0 to 50°C; or trifluoroacetic acid and anisole, phenol or triethylsilane at about -40°C to room temperature; allyl palladium(O) catalyzed transalkylation reaction in the presence of sodium or potassium salt of 2-ethyl hexanoic acid, see for example J. Org. Chem. 1982, 47, 587.

trimethylsilyl by hydrolysis or alcoholysis at room temperature.

In order to manufacture a readily hydrolysable ester of the carboxylic acids of formula I in accordance with embodiment a carboxylic acid of formula I is preferably reacted with a corresponding halide, preferably an iodide, containing the desired ester group. The reaction can be accelerated with the aid of a base such as an alkali metal hydroxide, an alkali metal carbonate or an organic amine such as triethylamine. The esterification is S. preferably carried out in an inert organic solvent such as dimethyl-acetamide, 0 15 hexamethylphosphoric acid triamide, dimethyl sulfoxide or, especially, dimethylformamide. The reaction is preferably carried out at a temperature in the range of about 0-40°C.

The manufacture of the salts and hydrates of the compounds of formula I or the hydrates of said salts in accordance with embodiment can be carried 20 out in a manner known per se; for example, by reacting a carboxylic acid of formula I or a salt thereof with an equivalent amount of the desired base, conveniently in a solvent such as water or an organic solvent ethanol, methanol, acetone and the like). Correspondingly, salt formation is brought about by the addition of an organic or inorganic salt or an acid. The 25 temperature at which the salt formation is carried out is not critical. The salt formation is generally carried out at room temperature, but it can be carried out at a temperature slightly above or below room temperature, for example in the range of 0OC to The manufacture of the hydrates usually takes place automatically in the course of the manufacturing process or as a result of the hygroscopic properties of an initially anhydrous product. For the controlled manufacture of a hydrate, a completely or partially anhydrous carboxylic acid of formula I or salt thereof can be exposed to a moist atmosphere at about +10*C to -23- Exemplary of the process for obtaining products in accordance with the invention are the following reaction schemes 1 and 2 below.

Scheme 1, embodiment (a)

R

1 -(X)s(CR 4 R 5 )m R HNS 0Y

(CH

2

N

1 C -R 2 00R

H

4 R 5 N S (CH 2 )n 0

N

wherein the symbols are as defined above.

:Scheme 2, embodiment (b) 0 5)n H

N-R

2 Br or C1I

NR

.0000. 0 0R 0 CQ 2 R9 0

H

R'(X)s-(0R 4

R

5 )m

(CH

2 )n N-R 2 000.R 0 wherein X is 0, S, NH and XY accordingly OH or SH or S- or NH 2 and the remaining substituent are as defined above.

-24- Examples Example 1 [(4-Hydroxy-phenylcarbamoyl)-methyll -pyridin- 1-ium-4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyll -8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino] -5-thia- 1-aza-bicyclo[4. 2.0] oct-2-ene-2-carboxylate H 0

NNIO

*S

C1 cc 0 To a solution of 68.4 mg (0.25 mmol) (2,4,5-trichloro-phenylsulfanyl)-acetic acid in 3 nml N,N-dimethylacetamide were added under stirring and Argon atmosphere 40.9 mg (0.25 mmol) 1,1'-carbonyldiimidazole. After 30 min, 136.4 10 mg (0.21 minol) (E)-(6R,7R)-7-amino-3- [1-11-[(4-hydroxy-phenylcarbamoyl)methyl] -pyridin- 1-ium-4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyll thia-1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylate trifluoro acetate was added in a single portion. After 3h the reaction mixture was poured on diethyl ether. The solid material was collected by filtration and triturated with ethyl acetate.

Yield: 112.0 mg beige powder IR(KCBr): 1770, 1678, 1642 cm-1 MS(ISP): 790.2 According to the procedure in Example 1 the following additional compounds were prepared: Example 2 [1-11-[(4-Hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1-ium-4ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyll -8-oxo-7- 1-benzothiazol-2ylsulfanyl)-acetylamino] -5-thia- 1-aza-bicyclo oct-2-ene-2-carboxylate 4 N 0

H

QN#~r~>0 With 70.0 mg (0.43 mmol) 1,1'-carbonyldiimidazole, 96.0 mg (0.43 mimol) (benzothiazol-2-ylsulfanyl)-acetic acid and 233.8 mg (0.36 mmol) amino-3- [1-[(4-hydroxy-phenylcarbamoyl).-methyl] -pyridin- 1-ium-4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyll-8-oxo-5-thia- 1-aza-bicyclo oct- 2-ene-2-carboxylate trifluoroacetate, in 4 nml N,N-dimethylacetamide.

Yield: 92.0 mg yellow powder IR(KBr): 1769, 1679, 1643, 1625 cm-1 MS(ISP): 743.3 Example 3 [2-(Benzothiazol-2-ylsulfanyl)-acetylamino 1-cyclopropylmethyl-2-oxo-pyrrolidin-3-ylidenemethyl)-8-oxo-5-thia- 1-aza-bicyclo oct- 2-ene-2-carboxylic acid

H

N N s x N With 220.0 mg (1.35 mmol) 1,1'-carbonyldiimidazole, 304.1 mg (1.35 mmnol) benothazo-2ylslfayl)aceicacid and 394.8 mg (1.13 mmol) amino-3-( 1-cyclopropylmethyl-2-oxo-pyrrolidfin-3-ylidenemethyl)-8-oxo-5-thia- 1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid in 7 ml N,N-dimethylformamide.

Yield: 173.0 mg orange powder IR(KBr): 1772, 1665, 1623 cm-1 MS(ISP): 557.1 Example 4 [1-[1-[(4-Hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1-ium-4ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyll -8-oxo-7- [2-(pyridin-4-ylsulfanyl)acetylamino] -5-thia-1-aza-bicyclo [4.2.01 oct-2-ene-2-carboxylate

HO

02 0 With 70.0 mg (0.43 nunol) 1,1'-carbonyldiimidazole, 72.8 mg (0.43 nmol) (pyridin-4-ylsulfanyl)-acetic acid and 232.8 mg (0.36 rnmol) 26amnino-3- [1-[(4-hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1-ium-4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyll-8-oxo-5-thia- 1-aza-bicyclo oct- 2-ene-2-carboxylate trifluoroacetate in 4 ml mal N,N-dimethylacetamide. The brown solid was purified by column chromatography on MCI gel (75-150g~, Mitsubishi Kasei Corporation) with a gradient of water: acetonitrile, 4:1, 3:1, 2: 1, 1: The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 58.0 mg beige lyophilisate IR(KBr): 1772, 1670, 1625 cm1 MS(ISP) 687.3 Example [1-11-[(3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1ium-4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyll [2-(naphthalen-2-ylsulfanyl)-acetylamino] -8-oxo-5-thia- 1-aza-bicyclo oct-2-ene-2-carboxylate 00 HQ 0*

HH

o N0 *00* 02- 0 With 58.3 mg (0.36 mmol) 1,1'-carbonyldiimidazole, 78.5 mg (0.36 mmol) (naphthalen-2-ylsulfanyl)-acetic acid and 200.0 mg (0.30 minol) 00 amino-3- [1-11- 13-fluoro-4-hydroxy-phenylcarbamoyl)-methyl] pyridin- 1-ium- *000~ 4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-5-thia-l1-aza-bicyclo- 14.2.0] oct-2-ene-2-carboxylate, trifluoroacetate in 4 ml N,N-dimethyl acetamide.

The resulting solid was purified by colun chromatography on MCI gel 150gi, Mitsubishi Kasei Corporation) with a gradient of water: acetonitrile 0, 4:1, 3:1, 2: 1, 1: The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 55.0 mg beige lyophilisate IR(KBr): 1770, 1680, 1650, 1628 cm'1 MS(ISP) 754.3 Example 6 [1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino] -5-thia- 1-aza-bicyclo oct- 2-ene-2-carboxylic acid -27- H

OH

CI

C0 2 H 0 With 146.0 mg (0.90 mmol) 1,1'-carbonyldiirmidazole, 244.4 mg (0.90 mmol) (2,4,5-trichloro-phenylsulfanyl)-acetic acid and 301.0 mg (0.73 inmol) (6R,7R)-7-amino-3- [1-(3-hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl] -8oxo-5-thia-1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid trifluoroacetate in ml N,N-dimethylacetamnide.

Yield: 180.0 mg beige powder IR(KBr): 1767, 1664, 1614 cm'1 MS(ISP): 654.1 00 10 Example 7 [1-(3-Hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-7- 0 [2-naphthalen-2-ylsulfanyl)-acetylaminol-5-thia- 1-aza-bicyclo [4.2.01 oct-2-ene- 2-carboxylic acid *ooN

OH

15 With 146.0 mg (0.90 mmol) 1,1'-carbonyldiimidazole, 196.5 mg (0.90 mmol) 0:0. (naphthalen-2-ylsulfanyl)-acetic acid and 301.0 mg (0.73 mmol) amino-3- [1-(3-hydroxy-benzyl)-2-oxo-pyrrolidin-3-ylidenemethyl] 1-aza-bicyclo[4.2.0]oct-2-ene-2-carboxylic acid trifluoroacetate in 5 ml N,Ndimethylacetamide. The resulting solid was purified by column chromatography on MCI gel (75-150g, Mitsubishi Kasei Corporation) with a gradient of water: acetonitrile 0, 4:1, 3:1, 2: 1, 1: The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 65.0 mg beige lyophilisate IR(KBr): 1771, 1663, 1589 cnf' MS(ISP): 602.2 Example 8 Mixture of (E)-(6R,7R)-8-oxo-3- and bipyrrolidinyl-3ylidenemethyl] -8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino] 1-aza-bicyclo oct-2-ene-2-carboxylic acid hydrochloride 28- Step a: Mixture of and -[(S)-1'-allyloxycarbonyl-2-oxobipyrrolidinyl-3-ylidenemethyll -8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino] -5-thia- 1-aza-bicyclo [4.2 .0]oct-2-ene-2-carboxylic acid C1

H

C1 0 0 C0 2 H 0 With 115.2 mg (0.71 mmol) 1,1'-carbonyldiirnidazole, 193.4 mg (0.71 mmol) (2,4,5-trichloro-phenylsulfanyl)-acetic acid and 329.1 mg (0.59 nunol) of a mixture of and 1'-allyloxycarbonyl-2-oxo- hi- 10 pyrrolidin-3-ylidenemethyl)-7-amino-8-oxo-5-thia- 1-aza-bicyclo oct-2-ene- 102-carboxylic acid trifluoroacetate in 6 mnl N,N-dimethylacetamide Prepared according to example Al.

Yield: 220.0 mg beige powder IR(KBr): 1774, 1678, 1624 cm1 MS(ISP): 703.2 Step b: Mixture of and -[(S)-2-oxo-I1,3']bipyrrolidinyl- 3-ylidenemethyl] -8-oxo-7- [2-(2,4,5-trichloro-phenylsulfanyl)-acetylamino] thia- 1-aza-bicyclo [4.2.01 oct-2-ene-2-carboxylic acid hydrochloride The product prepared in step a (220.0 mg, 0.31 mmol) was suspended in 12 ml dichioromethane and 124 g1 (0.50 mmol) N,O-bis-(trimethylsilyl)-acetamide was added. After a clear solution had formed, 5.60 mg (8.56 jimol) palladiumbis-(triphenylphosphine)-dichloride, 0.36 ml (6.30 mmol) acetic acid and 0.8 ml mmol) tributyltinhydride were added. After 45 min 40 jgd water was added to the suspension and the reaction mixture was poured under stirring on 200 mlA diethyl ether, containing 2 ml of a hydrochloric acid-saturated diethyl ether solution. The solid was collected by filtration and triturated with 40 ml ethyl acetate.

-29- Yield: 180.0 mg beige powder IR(KBr): 1771, 1661, 1582 cm1 MS(ISP): 619.1 Example 9 [1-[3-Fluoro-4-hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1ium-4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-7-(2-phenylsulfanyl- 1-aza-bicyclo oct-2-ene-2-carboxylate H r_6 O C0 2 000~ With 175.0 mg (1.08 mmol) 1,1'-carbonyldiimidazole, 182.0 mg (1.08 mnmol) 2- 10 (phenylthio)acetic acid and 500.0 mg (0.75 mmol) (E)-(6R,7R)-7-amino-3-[1-11- (3-fluoro-4-hydroxy-phenylcarbamoyl)-methyl pyridin-1-ium-4-ylmethyl] -2- 0 0oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-5-thia- 1-aza-bicyclo oct-2-ene-2carboxylate trifluoroacetate in 4 ml N,N-diinethylacetamide. The resulting solid was purified by colurnm chromatography on MCI gel (75-150g., Mitsubishi Kasei Corporation) with a gradient of water acetonitrile 4:1, 3:1, 2:1).

The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

o o o eYield: 90.0 mg beige lyophilisate IR(KBr): 1772, 1680, 1648 cm'1 20 MS(ISP): 704.4 (M 00 Example [2-(2,5-Dichloro-phenylsulfanyl)-acetylamino [1-[(3-fluoro-4hydroxy-phenylcarbamoyl)-methyl] -pyridin- 1-ium-4-ylmethyll -2-oxopyrrolidin-3-ylidenemethyl] -8-oxo-5-thia-1-aza-bicyclo 14.2.0] oct-2-ene-2carboxylate 00 coo- 0 With 72.9 mg (0.45 mnmol) 1,1'-cairbonyldiimidazole, 106.5 mg (0.45 mnmol) dichloro-phenylsulfanyl)acetic acid and 250.0 mg (0.37 mmol) amino-3- [(3-fluoro-4-hydroxy-phenylcarbamoyl)-methyl] pyridin- 1-ium- 4-ylmethyl] -2-oxo-pyrrolidin-3-ylidenemethyl] -8-oxo-5-thia- 1-azabicyclo oct-2-ene-2-carboxylate trifluoroacetate in 4 ml N,N-dimethyl acetamide. The resulting solid was purified by column chromatography on MCI gel (75- 150gj, Mitsubishi Kasei Corporation) with a gradient of water: acetonitrile (1 0, 4: 1, 3 1, 2 1, 1 The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 74.5 mg beige lyophilisate IR(KBr): 1772, 1680, 1650 cm- 1 MS(ISP): 772.3 (M+H Example 11 (E)-(6R,7R)-8-Oxo-3- [1 ,3']bipyrrolidinyl-3-ylidenemethyll phenylsulfanyl-acetylamino)-5-thia-1-aza-bicyclo oct-2-ene-2- **:carboxylicacid hydrochloride 1)

H

NH

15 C0 2 H 0 1'-allyloxycarbonyl-2-oxo- bipyrrolidinyl-3ylidenemethyl] -8-oxo-7-(2-phenylsulfanyl-acetylamino)-5-thia- 1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid (370.0 mg, 0.62 mmol) was dissolved in 20 ml dichloromethane and treated successively with bis(triphenylphosphine)palladium(II) chloride (10.9 mg, 0.015 mmol), acetic acid (0.71 ml, 12.4 mmol) and tributyltin hydride (1.67 ml, 6.20 minol). After min, the suspension was poured on 250 ml diethyl ether containing 3 ml of a hydrochloric acid-saturated diethyl ether solution and stirred for 1h. The suspension was filtered, the solid material was triturated with ethyl acetate for 1h and dried in high vacuum.

Yield: 25.5% beige solid MS(ISP): 515.3 IR(KBr): 1776, 1666, 1632 cm'1 Example 12 1-Cyclopropyl-2-oxo-pyrrolidin-3-ylidenemethyl)-7- [2- (naphthalen-2-ylsulfanyl)-acetylamino] -8-oxo-5-thia- 1-aza-bicyclo oct-2ene-2-carboxylic acid imidazole salt (1:1) -31- S i7 N.

C0 2 H 0 Yield: 42.6 beige solid MS(ISP): 536.3 IR(KiBr): 1769, 1664, 1624 cm'1 Example 13 [1-(4-Hydroxy-phenyl)-2-oxo-pyrrolidin-3-ylidenemethy] [2- (naphthalene-2-ylsulfanyl)-acetylamino] -8-oxo-5-thia- 1-aza-bicyclo oct-2ene-2-carboxylic acid sodium salt (1:1)

H

N S N\

OH

C0 2 Na 0 With 140.0 mg (0.86 mmol) 1,1'-carbonyldiimidazole, 187.7 mg (0.86 mmol) (2naphthylthio)acetic acid and 230.0 mg (0.60 mmol) (E)-(6R,7R)-7-amino-3-[1- (4-hydroxy-phenyl)-2-oxo-pyrrolidin-3-ylidenemethyl -8-oxo-5-thia- 1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid in 4 ml N,N'-dimethylacetamide. The resulting yellow solid was suspended in 3 ml water and converted into its sodium salt by treatment with 500 jil 1M NaOH. The solution was chromatographed on MCI gel with a gradient of water acetonitrile 9: 1).

The organic solvent was stripped off at a rotary evaporator and the aqueous phase was freeze-dried.

Yield: 46.0 yellow lyophilisate MS(ISP): 588.3 IR(Nujol): 1761, 1659, 1591 cm' Example 14 (E)-(6R,7R)-8-Oxo-3- [2-oxo- 1-(4-sulfamoyl-benzyl)-pyrrolidin-3-ylidenemethyl 7- 12-phenylsulfanyl)-acetylamino] -5-thia- 1-aza-bicyclo oct-2-ene-2carboxylic acid -32-

CO

2

H

Yield: 75.3 beige solid MS(ISN): 613.2 (M-H) IR(KBr): 1770, 1667, 1602 cm- 1 The following example illustrates pharmaceutical preparations containing an antimicrobial composition provided by the present invention: Example A Production of dry ampoules for parenteral administration: 10 A mixture of 0,25-8 g of vinyl-pyrrolidinone cephalosporin derivative and 0,25- 8 g carbapenem antibiotic or 0,25-8 g p-lactamase inhibitor is prepared in the usual manner and filled into an ampoule as a) mixtures of dry powders (crystalline, amorpheous or lyophilisates powder) of vinyl-pyrrolidinone cephalosporin derivative and carbapenem antibiotic or 15 P-lactamase inhibitor; or b) lyophilisates of the mixed solutions of vinyl-pyrrolidinone cephalosporin derivative and carbapenem antibiotic or -lactamase inhibitor.

a...i The dry powders (crystalline, amorpheous or lyophilisates powder) of vinylpyrrolidinone cephalosporin derivative and carbapenem antibiotic or Plactamase inhibitor can be filled in separate ampoules and mixed prior to the administration.

Claims (15)

1. Antimicrobial composition comprising a combination of a first ingredient selected from vinyl-pyrrolidinone cephalosporin derivatives of the general formula I R 4 H 0 3, nN-R2 0 0 R 3 wherein R 1 is lower alkyl, halogen, phenyl, benzyl, styryl, naphthyl or heterocyclyl; the lower alkyl, phenyl, benzyl, styryl, naphthyl and heterocyclyl being optionally substituted by at least one of halogen, hydroxy, optionally substituted lower alkyl, optionally substituted lower alkoxy, optionally substituted phenyl, amino, lower alkylamino, di-lower alkylamino, carboxy, lower alkyl-carboxy, carbamoyl or lower alkylcarbamoyl; R 4 R 5 independently are hydrogen, lower alkyl or phenyl; X is S, O, NH or CH 2 n is 0, 1 or 2; m is 0 or 1; s is 0 or 1; R 2 is hydrogen, hydroxy, carbamoylmethyl, -CH 2 -CO-NH-R 6 lower alkyl-Qr, cycloalkyl-Qr, lower alkoxy, lower alkenyl, cycloalkenyl-Qr, lower alkynyl, aralkyl-Qr, aryl-Qr, aryloxy, aralkoxy, heterocyclyl or heterocyclyl-Qr; the lower alkyl-, cycloalkyl-, lower alkoxy-, lower alkenyl-, cycloalkenyl-, lower alkynyl-, aralkyl-, aryl-, aryloxy-, aralkoxy- and the S heterocyclyl-group may be substituted with at least one group selected from carboxy, amino, nitro, cyano, S0 2 -NH-R 6 optionally fluoro substituted lower alkyl, lower alkoxy, hydroxy, halogen, CONR 6 R 7 -CH 2 -CONR 6 R 7 N(R 7 )COOR 8 R 7 CO-, R 7 0CO-, R7COO-, C(R 7 R 9 )CO 2 R 8 -C(R 7 R 9 )CONR 7 R 10 wherein R 6 is hydrogen, lower alkyl, cycloalkyl or aryl; R 7 and R 9 are independently hydrogen or lower alkyl; R 8 is hydrogen, lower alkyl, lower alkenyl or a carboxylic acid protecting group; and R 1 0 is hydrogen, w-hydroxy-alkyl, phenyl, naphthyl or heterocyclyl; the phenyl, naphthyl or heterocyclyl being unsubstituted or substituted with at least one of the groups of optionally protected hydroxy, halogen, optionally substituted lower alkyl or w-hydroxyalkyl, optionally substituted lower alkoxy and/or cyano; or R 7 and R10 form together a group of formula Q is -CH 2 -CH-lower-alkyl, -CO- or SO 2 r is 0 or 1; and R 3 is hydroxy, or -OM and M represents an alkali metal; as well as readily hydrolysable esters thereof, pharmaceutically acceptable salts of said compounds and hydrates of the compounds of formula I and of their esters and salts; and C04078 a second ingredient selected from the class of carbapenem antibiotics and pharmaceutically acceptable salts thereof or from p-lactamase inhibitors and pharmaceutically acceptable salts thereof.

2. Antimicrobial composition according to claim 1, comprising a first ingredient selected from vinyl-pyrrolidinone cephalosporin derivatives of the general formula I wherein R 1 is phenyl, 2-naphthyl, 2,5-dichlorophenyl, 2,4,5-trichlorophenyl, 2-benzothiazolyl, 4-pyridinyl; R 2 is cyclopropyl, cyclopropylmethyl-, 3- or 4-hydroxybenzyl, 4-hydroxyphenyl, 4-sulfamoylbenzyl, pyrrolidin-3-yl, 1-[N-(4-hydroxy)phenyl]-carbamoylmethyl-pyridin-1-ium-4-yl, fluoro-4-hydroxy)phenyl]-carbamoylmethyl-pyridin-l-ium-4-yl; R 3 is hydroxy, lower-alkoxy, or -OM and M represents an alkali metal; X is S; (CR 4 R 5 )m is -CH 2 and pharmaceutically acceptable salts thereof.

3. Antimicrobial composition according to claim 1 or 2, comprising a combination of a first ingredient selected from vinyl-pyrrolidinone cephalosporin o 15 derivatives of the general formula I as defined in claim 1 and pharmaceutically acceptable salts thereof; and a second ingredient selected from carbapenem antibiotics and pharmaceutically acceptable salts thereof.

4. Antimicrobial composition according to claim 3, wherein the carbapenem S" antibiotic is imipenem. S 20

5. Antimicrobial composition according to claim 1 or 2, comprising a combination of a first ingredient selected from vinyl-pyrrolidinone cephalosporin derivatives of the general formula I as defined in claim 1 and pharmaceutically acceptable salts thereof; and a second ingredient selected from p-lactamase inhibitors and pharmaceutically acceptable salts thereof.

6. Antimicrobial composition according to claim 5, wherein the 13- lactamase inhibitor is (Z)-(2S,3S,5R)-3-(2-cyanoethenyl)-3-methyl-4,4,7-trioxo-4- thia-1-aza-bicyclo[3.2.0]heptane-2-carboxylic acid.

7. Antimicrobial composition according to any one of claims 1 to 6, wherein the ratio by weight of the first ingredient to the second ingredient is about 1:20 to about 20:1.

8. An antimicrobial composition, substantially as hereinbefore described with reference to any one of the examples.

9. A pharmaceutical preparation containing an antimicrobial composition according to any one of claims 1 to 8 and a therapeutically inert carrier.

10. A method for the treatment or prophylaxis of infectious diseases in a mammal requiring said treatment or prophylaxis, which method includes or consists of administering to said mammal an effective amount of at least one antimicrobial composition according to any one of claims 1 to 8, or of a pharmaceutical preparation according to claim 9. C04078

11. An antimicrobial composition according to any one of claims 1 to 8 or a pharmaceutical preparation according to claim 9 when used in the treatment or prophylaxis of infectious diseases.

12. The use of an antimicrobial composition according to any one of claims 1 to 8 for the manufacture of a medicament for the treatment or prophylaxis of infectious diseases.

13. An antimicrobial composition or pharmaceutical preparation when used as claimed in claim 11 wherein the infectious diseases include MRSA infections.

14. The use of an antimicrobial composition as claimed in claim 12, wherein the infectious diseases include MRSA infections. The method according to claim 10 wherein the infectious diseases include MRSA infections. Dated 21 February, 2002 F.Hoffmann-La Roche AG

15 Patent Attorneys for the Applicant/Nominated Person SPRUSON FERGUSON **S S* So g *oot ot f f ft ft o f [R:LIBW]45695.doc:SOH